Understanding stochastic and deterministic assembly processes in microbial communities along temporal, spatial and environmental scales.

Identifying the main drivers of community assembly remains an open fundamental question in ecology. Dispersal processes introduce randomness in community composition while selection for particular environments creates predictable assemblages. However, the interaction between selection and dispersal processes is still poorly understood. Here, we address this question in bacterial and microeukaryotic communities inhabiting a highly dynamic system of ephemeral (hyper)saline lakes. We show that the combination of beta-diversity decomposition methods and a temporal approach based on colonization and extinction dynamics yields new insights into the relative effect of selection and dispersal along environmental gradients. Selective pressure and dispersal-related processes simultaneously shape each local community with variable strength and effect. The dominance of selection vs. dispersal shifted from stochastic to deterministic assembly as salinity increased along the gradient. This transition also had an impact on the temporal dynamics of the lakes as community turnover decreased at high salinities because both colonization and extinction rates slowed down. Only microeukaryotic richness decreased along the gradient due to lower effective colonization at higher salinities, suggesting that the net effect of selection and dispersal is determined by both environmental conditions and the idiosyncrasy of the different microbial ecologies. Our results emphasize the use of temporal approaches in combination with standard statistical methods for a better understanding of the dynamic processes underlying community assembly.

Absence of stress-promoted facilitation coupled with a competition decrease in the microbiome of ephemeral saline lakes.

Salinity fluctuations constitute a well-known high stress factor strongly shaping global biological distributions and abundances. However, there is a knowledge gap regarding how increasing saline stress affects microbial biological interactions. We applied the combination of a probabilistic method for estimating significant co-occurrences/exclusions and a conceptual framework for filtering out associations potentially linked to environmental and/or spatial factors, in a series of connected ephemeral (hyper) saline lakes. We carried out a network analysis over the full aquatic microbiome-bacteria, eukarya, and archaea-under severe salinity fluctuations. Most of the observed co-occurrences/exclusions were potentially explained by environmental niche and/or dispersal limitation. Co-occurrences assigned to potential biological interactions remained stable, suggesting that the salt gradient was not promoting interspecific facilitation processes. Conversely, co-exclusions assigned to potential biological interactions decreased along the gradient both in number and network complexity, pointing to a decrease of interspecies competition as salinity increased. Overall, higher saline stress reduced microbial co-exclusions while co-occurrences remained stable suggesting decreasing competition coupled with lack of stress-gradient promoted facilitation in the microbiome of ephemeral saline lakes.

Ecological and Metabolic Thresholds in the Bacterial, Protist, and Fungal Microbiome of Ephemeral Saline Lakes (Monegros Desert, Spain).

We studied the 16S and 18S rRNA genes of the bacterial, protist, and fungal microbiomes of 131 samples collected in 14 ephemeral small inland lakes located in the endorheic area of the Monegros Desert (NE Spain). The sampling covered different temporal flooding/desiccation cycles that created natural salinity gradients between 0.1% (w/v) and salt saturation. We aimed to test the hypothesis of a lack of competitive advantage for microorganisms using the "salt-in" strategy in highly fluctuating hypersaline environments where temperature and salinity transitions widely vary within short time periods, as in ephemeral inland lakes. Overall, 5653 bacterial zOTUs and 2658 eukaryal zOTUs were detected heterogeneously distributed with significant variations on taxonomy and general energy-yielding metabolisms and trophic strategies along the gradient. We observed a more diverse bacterial assembly than initially expected at extreme salinities and a lack of dominance of a few "salt-in" organisms. Microbial thresholds were unveiled for these highly fluctuating hypersaline environments with high selective pressures. We conclude that the extremely high dynamism observed in the ephemeral lakes of Monegros may have given a competitive advantage for more versatile ("salt-out") organisms compared to those better adapted to stable high salinities usually more common in solar salterns. Ephemeral inland saline lakes offered a well-suited natural framework for highly detailed evolutionary and ecological studies.

Dynamics and ecological distributions of the Archaea microbiome from inland saline lakes (Monegros Desert, Spain).

We characterized the rich Archaea microbiome of shallow inland lakes (Monegros Desert, NE Spain) by 16S rRNA gene tag sequencing covering a wide salinity range (0.1%-40% w/v) along 3 years. Up to 990 operational taxonomic units (OTUs; >97% identity) were detected allocated in 14 major archaeal phyla and heterogeneously distributed along the salt gradient. Dynamics and idiosyncratic ecological distributions were uncovered for the different phyla. A high genetic richness was observed for Woesearchaeota and Pacearchaeota (>370 OTUs each), followed by Halobacteria (105), Nanohaloarchaeota (62) and Thermoplasmata (19). Overall, the distribution of genetic richness was strongly correlated with environmental niche amplitude, but not with occurrence. We unveiled high occurrence for a very rich Woesearchaeota assemblage, and an unexpected positive correlation of Pacearchaeota abundance with salinity at >15% dissolved salt content. The estimated dynamic behaviour (temporal 'turnover' rates of presence/absence data) unveiled Thaumarchaeota and Halobacteria as the most dynamic groups, and Aenigmarchaeota and Thermoplasmata as the most stable. The DPANN Pacearchaeota, Woesearchaeota, and Nanohaloarchaeota showed intermediate rates, suggesting higher resilience to environmental perturbations. A rich and dynamic Archaea microbiome was unveiled, including unseen ecological traits for relevant members of the still largely unknown DPANN group, supporting a strong ecological differentiation between Pacearchaeota and Woesearchaeota.

A Randomized Trait Community Clustering approach to unveil consistent environmental thresholds in community assembly.

Similarities and differences of phenotypes within local co-occurring species hold the key to inferring the contribution of stochastic or deterministic processes in community assembly. Developing both phylogenetic-based and trait-based quantitative methods to unravel these processes is a major aim in community ecology. We developed a trait-based approach that: (i) assesses if a community trait clustering pattern is related to increasing environmental constraints along a gradient; and (ii) determines quantitative thresholds for an environmental variable along a gradient to interpret changes in prevailing community assembly drivers. We used a regional set of natural shallow saline ponds covering a wide salinity gradient (0.1-40% w/v). We identify a consistent discrete salinity threshold (ca. 5%) for microbial community assembly drivers. Above 5% salinity a strong environmental filtering prevailed as an assembly force, whereas a combination of biotic and abiotic factors dominated at lower salinities. This method provides a conceptual approach to identify consistent environmental thresholds in community assembly and enables quantitative predictions for the ecological impact of environmental changes.

Microbial composition, potential functional roles and genetic novelty in gypsum-rich and hypersaline soils of Monegros and Gallocanta (Spain).

Soil microbial communities (both Bacteria and Archaea) were studied after 16S rRNA genes massive sequencing in two hypersaline and gypsum-rich contrasted sites located in NE Spain. Soil microbial communities were also locally analysed according to environmental variables, including geological, physico-chemical, biogeochemically, and climatic data. Typical soil characteristics, climate data, and plant composition clearly split the two sites and major differences among the microbial communities for the areas were initially expected. Overall, high values of microbial species richness (up to 2300 taxa) and ecological diversity was detected in both sites. High genetic novelty levels were found mostly to environmental sequences, highlighting the high potential for microbiological studies. In contrast to the initial expectations, a substantial overlapping between Monegros and Gallocanta microbes was observed, indicating a high similarity despite of the geographical, botanical and environmental distances between sites, in agreement with both high dispersal and local selection inherent to the microbial world. The potential biogeochemical cycling showed small differences between sites, with presence of photosynthetic green and purple sulfur bacteria, cyanobacteria and aerobic and anaerobic chemolitotrophs. Potential for aerobic methane oxidation and anaerobic methanogenesis was observed in both sites, with predominance of potential nitrification mostly by ammonia-oxidizing archaea, nitrite oxidation and denitrification, and minor contribution for nitrate reduction and nitrate ammonification. The predicted functions based on the taxonomic composition showed high overlapping between the two studied regions, despite their difference in gypsum richness.